Centrosomes and BRCA1

中心体和 BRCA1

• 批准号: 7395006
• 负责人: JEFFREY D PARVIN
• 金额: $ 23.27万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7395006
• 关键词: BRCA1 Protein; BRCA1 gene; Biological; Biological Assay; Breast; Cell Line; Cells; Centrosome; Chimeric Proteins; DNA; Data; Epithelial Cells; Epithelium; Etiology; Event; Growth; In Vitro; Laboratories; Learning; Lesion; Lysine; Malignant neoplasm of ovary; Mammary Gland Parenchyma; Mammary gland; Mass Spectrum Analysis; Microtubules; Modification; Mutation; Normal Cell; Numbers; Organelles; Pan Genus; Pathway interactions; Phenotype; Process; Protein Binding; Protein Inhibition; Proteins; Publishing; Regulation; Research; Research Personnel; Small Interfering RNA; Testing; Tubulin; Tumor Suppressor Proteins; Ubiquitination; Work; cell type; in vitro Assay; in vivo; malignant breast neoplasm; mutant; osteosarcoma; polypeptide; programs; research study; tissue culture; tumor; ubiquitin ligase; ubiquitin-protein ligase

Why mutations in the BRCA1 gene result in breast and ovarian cancer is unknown. Recent data from this laboratory suggest that the ubiquitination activity of the BRCA1 tumor suppressor regulates the duplication and function of centrosomes specifically in mammary epithelial cells in tissue culture. Using a transient assay to inhibit BRCA1 function, we have not observed centrosome amplification in cell lines derived from non- breast tissue. Centrosome amplification has been observed in the earliest and most aggressive breast cancer lesions, and it is likely that this function of BRCA1 is critical in the etiology of these tumors. Our data has revealed a new biological pathway, which controls centrosome number and function and which depends on the function of BRCA1 in breast cells. This project will dissect the BRCA1 function via three aims. 1.) Polypeptide targets of BRCA1 ubiquitination will be identified from centrosomes purified from breast cell lines. Proteins modified by the BRCA1-dependent ubiquitination activity will be assayed for effects on centrosome duplication. 2.) The effects of BRCA1-dependent ubiquitination on centrosome microtubule nucleation function will be assayed in vitro and in vivo, and the proteins and ubiquitinated substrates identified in aim 1 will also be tested for modulating the function of BRCA1 on this organelle. 3.) We will identify the factors in non-breast cells which render the BRCA1 ubiquitination activity redundant, and test whether the newly identified factor plus BRCA1 regulate centrosome amplification in these cell types. This project will identify the mechanism by which BRCA1 regulates centrosome duplication and centrosome function in breast cells. One of the earliest changes in the cells in an incipient breast cancer is a problem with the cellular machine that segregates the DNA when the breast cell divides. Research in this laboratory has found that the function of this machine, called a centrosome, is controlled in breast cells by the BRCA1 protein. This project will dissect the workings of the centrosome and determine how BRCA1 controls its function in normal cells, and we will learn how this process is changed when BRCA1 function is lost by mutation as happens in breast cancer.

为什么 BRCA1 基因突变会导致乳腺癌和卵巢癌尚不清楚。最新数据来自于此 实验室表明 BRCA1 肿瘤抑制因子的泛素化活性可调节复制 和中心体的功能，特别是组织培养中乳腺上皮细胞的功能。使用瞬态测定 为了抑制 BRCA1 功能，我们尚未观察到来自非非 BRCA1 细胞系的中心体扩增。 乳房组织。在最早和最具攻击性的乳房中观察到中心体扩增 癌症病变，并且 BRCA1 的这种功能很可能在这些肿瘤的病因学中至关重要。我们的数据 揭示了一种新的生物途径，它控制中心体的数量和功能，并且取决于 BRCA1 在乳腺细胞中的功能。该项目将通过三个目标剖析 BRCA1 功能。 1.) BRCA1 泛素化的多肽靶标将从乳腺细胞纯化的中心体中鉴定出来 线。将检测由 BRCA1 依赖性泛素化活性修饰的蛋白质对 中心体复制。 2.) BRCA1依赖性泛素化对中心体微管的影响 将在体外和体内测定成核功能，以及蛋白质和泛素化底物 目标 1 中确定的 BRCA1 在此细胞器上的功能调节也将进行测试。 3.) 我们会 确定非乳腺细胞中导致 BRCA1 泛素化活性冗余的因素，并测试 新发现的因子加上 BRCA1 是否可以调节这些细胞类型中的中心体扩增。这 项目将确定 BRCA1 调节中心体复制和中心体的机制 在乳腺细胞中发挥作用。 早期乳腺癌细胞中最早的变化之一是细胞机器的问题 当乳腺细胞分裂时，它会分离 DNA。本实验室的研究发现， 这台机器的功能称为中心体，在乳腺细胞中由 BRCA1 蛋白控制。这 该项目将剖析中心体的运作并确定 BRCA1 在正常情况下如何控制其功能 细胞，我们将了解当 BRCA1 功能因突变而丧失时，这个过程是如何改变的，就像在 乳腺癌。